Unit

UNIT 21.2 Separation of Histone Variants and Post-Translationally Modified Isoforms by Triton/Acetic Acid/Urea Polyacrylamide Gel Electrophoresis

  1. Colleen A. Ryan,
  2. Anthony T. Annunziato

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb2102s45

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Ryan, C. A. and Annunziato, A. T. 2001. Separation of Histone Variants and Post-Translationally Modified Isoforms by Triton/Acetic Acid/Urea Polyacrylamide Gel Electrophoresis. Current Protocols in Molecular Biology. 45:21.2:21.2.1–21.2.10.

Author Information

  1. Boston College, Chestnut Hill, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1999

Abstract

Due to their similarities in size and charge, complete resolution of histones by electrophoresis poses a considerable challenge. The addition of nonionic detergents to the traditional acetic acid/urea (AU) polyacrylamide gel electrophoresis (PAGE) system has afforded an excellent method to separate not only the different modified forms of histones, but also the primary sequence variant subtypes of selected histone species; it is widely used to separate histones with varying levels of acetylation. This unit describes the use of gels containing the nonionic detergent Triton X-100, referred to as Triton/acetic acid/urea (TAU) polyacrylamide gels, for analysis of histones. Also included are support protocols detailing several accessory techniques: assembly of gel plates for the TAU gel, preparation of histones from isolated nuclei in a solubilized form amenable to electrophoresis, and electrophoretic transfer of proteins from these gels to PVDF membranes.